Identification of key cell subpopulations and associated biomarkers for mouse dry eye disease based on the...

Dry eye disease (DED) is characterized by tear film instability and chronic inflammation, with the cornea serving as a key target site vulnerable to inflammatory damage. This study integrated single-cell and bulk transcriptomics combined with Scissor and CIBERSORTx algorithms to screen corneal key cell populations and biomarkers in murine DED. Corneal wing cells were identified as core disease-associated cells, among which interferon-response wing cells (IFN-response-wing) were the pivotal pathogenic subtype. Three IFN-response-wing signature biomarkers Upk3b, Dusp1 and Ly6e were screened, enriched in immune pathways including antigen processing and presentation, primary immunodeficiency, and cytokine receptor interaction. We constructed a chemical-induced DED mouse model for multi-layered validation via immunofluorescence, Real-time polymerase chain reaction (RT-qPCR) and Western blot. Immunofluorescence revealed tissue-specific protein patterns: DED corneal epithelium exhibited elevated IRF1, Dusp1 and Ly6e protein yet reduced Upk3b, with Ly6e co-localized with IRF1-positive cells. However, RT-qPCR and Western blot of whole corneal tissue demonstrated significant upregulation of Upk3b, Dusp1 and Ly6e at both mRNA and total protein levels. Collectively, IFN-response-wing drives DED pathogenesis, and Upk3b, Dusp1, Ly6e act as tissue-specific DED biomarkers with inconsistent trends between epithelial localized protein and global corneal molecular abundance, providing novel mechanistic clues for DED diagnosis and targeted therapy.